scholarly journals Expression levels of caspase-3 and gasdermin E and their involvement in the occurrence and prognosis of lung cancer

Author(s):  
yuanli huang ◽  
GuangHui Zhang ◽  
Qing Zhu ◽  
Xia Wu ◽  
LIGao Wu

Abstract Background Pyroptosis plays a dual role in the development of cancer and malignancy, and as such, may potentially be a new target for cancer treatment. However, the inflammatory response to pyroptosis may have adverse effects on the body. The roles of gasdermin E (GSDME), caspases, and related proteins associated with pyroptosis in cancer remain controversial. This study aimed to explore whether the expression levels of caspase-3 and GSDME affect the clinical stage, pathological grade, and survival prognosis of patients with lung cancer. Methods We examined the protein levels of GSDME, caspase-3, caspase-8, and caspase-9 in lung tissues from 100 patients with lung cancer by using immunohistochemistry. Results We found that GSDME, caspase-3, and caspase-8 were more highly expressed in the tumor tissues than in the adjacent normal tissues. Moreover, we found that GSDME could serve as a prognostic factor because there was a positive correlation between its expression level and the postoperative survival rate of patients with lung cancer. Conclusions GSDME may be an independent factor affecting the prognosis of patients with lung cancer. However, the role of GSDME and its related proteins in cancer requires further research.

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Xinlu Wang ◽  
Xuanxuan Hao ◽  
Youping Wang ◽  
Bin Li ◽  
Lin Cui ◽  
...  

Shenfuyixin granule (SFYXG, i.e., Xinshuaikang granule) is a prescription, commonly used in the clinical experience, which plays a significant role in the treatment of heart failure. The purpose of this present research was to investigate the protective effect of SFYXG, and the mechanism about anti-H2O2-induced oxidative stress and apoptosis in the neonatal rat cardiomyocytes. Myocardial cells, as is well known, were divided into 4 groups: normal, model, SFYXG, and coenzyme Q10 group, respectively. Cells viability was determined by MTT assay. Flow cytometry and AO/EB staining were implemented to test the apoptosis rate and intracellular reactive oxygen species (ROS) level. Mitochondrion membrane potential (MMP) was evaluated by JC-1 fluorescence probe method. The myocardial ultrastructure of mitochondrion was measured by electron microscope. The related mRNA expression levels of Bax, Bcl-2, Caspase-3, caspase-8, and caspase-9 were detected by real-time polymerase chain reaction (PCR). Also, the expression levels of Bax and Bcl-2 protein were detected by Western blot, and the expression levels of caspase-3, caspase-8, and caspase-9 protein were tested by caspase-Glo®3 Assay, caspase-Glo®8 Assay, and caspase-Glo®9 Assay, respectively. GAPDH was used as the internal reference gene/protein. The results revealed that SFYXG (0.5 mg/ml) raised the viability of myocardial cell, weakened the apoptosis rate and ROS level, corrected the mitochondrion membrane potential stability, and improved cell morphology and ultrastructure of myocardial mitochondrion. Furthermore, SFYXG upregulated the antiapoptosis gene of Bcl-2, but downregulated the proapoptosis genes of Bax, caspase-3, and caspase-9. In conclusion, SFYXG could appear to attenuate myocardial injury by its antioxidative and antiapoptosis effect.


2018 ◽  
Vol 38 (5) ◽  
Author(s):  
Yan-Wu Zhou ◽  
Rong Li ◽  
Chao-Jun Duan ◽  
Yang Gao ◽  
Yuan-Da Cheng ◽  
...  

Chromosome 14 ORF 166 (C14orf166), a protein involved in the regulation of RNA transcription and translation, has been reported to possess the potency to promote tumorigenesis; however, the role of C14orf166 in non-small-cell lung cancer (NSCLC) remains unknown. The purpose of the present study was to assess C14orf166 expression and its clinical significance in NSCLC. Immunohistochemical staining, quantitative real-time PCR (qRT-PCR), and Western blotting were used to detect the C14orf166 protein and mRNA expression levels in NSCLC tissues compared with adjacent normal tissues, as well as in NSCLC cells lines compared with normal human bronchial epithelial cells (HBE). Then, the correlations between the C14orf166 expression levels and the clinicopathological features of NSCLC were analyzed. Additionally, the Cox proportional hazard model was used to evaluate the prognostic significance of C14orf166. We found that C14orf166 expression increased in carcinoma tissues compared with their adjacent normal tissues at the protein (P<0.001) and mRNA levels (P<0.001). High expression of C14orf166 was significantly associated with the T stage (P=0.006), lymph node metastasis (P=0.001), advanced TNM stage (P<0.001), and chemotherapy (P<0.001). Moreover, according to the survival analysis, patients with overexpressed C14orf166 were inclined to experience a shorter overall survival and disease-free survival time (P<0.001). Multivariate COX analysis implied that C14orf166 was an independent prognostic biomarker. Taken together, our findings indicate that the overexpression of C14orf166 may contribute to the disease progression of NSCLC, represent a novel prognostic predictor and help high-risk patients make better decisions for subsequent therapy.


2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Xiaolin Xu ◽  
Shuqin Yu ◽  
Xiaoyuan Liu ◽  
Ying Feng

Objective. Ultrasound-targeted microbubble destruction (UTMD) technique has recently been developed as a nonviral delivery of gene therapy. This study aimed at investigating the survival and apoptosis of ovarian cancer cell line OVCA-433 by inhibiting Livin expression through ultrasound-targeted microbubble destruction. Methods. We synthesized a targeted microbubble agent for UTMD-mediated shRNA against Livin gene in human ovarian cancer OVCA-433 cells. Lipid microbubbles were conjugated with a luteinizing hormone-releasing hormone analog (LHRHa) by an avidin-biotin linkage to target the ovarian cancer OVCA-433 cells expressing LHRH receptors. The microbubbles were mixed with the recombinant plasmid harboring shRNA-Livin. shRNA-Livin was transfected into OVCA-433 cells upon exposure to 1 MHz pulsed ultrasound beam (0.5 W/cm2) for 8 s. Cell survival was measured by the MTT assay, cell apoptosis by flow cytometry using annexin V/PI double staining, and cell ultrastructure by using the transmission electron microscope. The mRNA and protein expression levels of caspase-3 and caspase-8 were detected by RT-qPCR and western blotting. Results. UTMD-mediated delivery of shRNA-Livin remarkably reduced the survival of OVCA-433 cells but promoted the apoptosis compared with shRNA-Livin alone, shRNA-Livin plus nontargeted microbubbles, and shRNA-Livin plus LHRHa-conjugated microbubbles containing shRNA-Livin with or without exposure to ultrasound pulses. It was also found that UTMD-mediated delivery of shRNA-Livin notably declined the mRNA and protein expression levels of caspase-3 and caspase-8 in OVCA-433 cells compared with shRNA-Livin alone, shRNA-Livin plus nontargeted microbubbles, and shRNA-Livin plus LHRHa-conjugated microbubbles containing shRNA-Livin with or without exposure to ultrasound pulses. Conclusion. Our experiment verifies the hypothesis that ultrasound mediation of targeted microbubbles can enhance the transfection efficiency of shRNA-Livin in ovarian cancer cells.


2021 ◽  
Vol 14 (12) ◽  
pp. 1834-1842
Author(s):  
Xin Hua ◽  
◽  
Guo-Sheng Gao ◽  
Xiao-Lei Ye ◽  
◽  
...  

AIM: To investigate the inhibitory effect of the combined use of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and oridonin on choroidal melanoma cell lines, and to explore its underlying mechanism. METHODS: MUM-2B and C918 cells were treated with different concentrations of TRAIL and oridonin, and MTT assay used to evaluate the inhibition rate of the two compounds on cells. Then, the cell cycle distribution and apoptosis were detected by flow cytometry, and changes in apoptosis-related proteins such as death receptor 5 (DR5), a-caspase-3, and x-linked inhibitor of apoptosis protein (XIAP) were detected by Western blot. MUM-2B cells were transfected with si-DR5, which interfered with the expression of the DR5 gene. MTT and Western blot assay were used to detect cell activity and apoptosis-related proteins. RESULTS: When TRAIL and oridonin were simultaneously administered to the MUM-2B cells, the apoptosis rate was significantly higher than that by the two drugs individually. However, the effect of combined use of TRAIL and oridonin on C918 cells was not significantly different from that used alone. Cell cycle analysis showed that TRAIL and oridonin could induce G2/M arrest in MUM-2B cells. The Western blot results showed that the protein expression levels of the DR5, a-caspase-3, and BAX increased, while the expression levels of the anti-apoptosis-related proteins XIAP and BCL-2 were suppressed when TRAIL and oridonin simultaneously administered to MUM-2B cells. Interfering the expression of DR5 gene in MUM-2B cells could reverse the inhibitory effect of oridonin and TRAIL on the proliferation and apoptosis induction of MUM-2B cells. CONCLUSION: The inhibitory effects of oridonin and TRAIL on MUM-2B cells are significantly enhanced when they were administered as a combined treatment, which may ascribe to up-regulation of DR5.


2019 ◽  
Author(s):  
Zhixiong Chen ◽  
Jing Wang ◽  
Anquan Yang ◽  
Lihua Zhang ◽  
Yaojia Lu ◽  
...  

Abstract Background: Previous studies demonstrated that pearl extract (PE) promotes wound healing and skin whitening. However, whether PE can inhibit ultraviolet (UV) photodamage in HaCaT cells remains unclear. In this study, an in vitro photoaging cell model was established to observe the effect of PE on UV-induced damage and apoptosis of HaCaT cells. The aim was to provide a reference for future development of natural sunscreen agents. Results: PE concentrations of 0.1 and 1 μg/mL were considered as the most effective and safe concentrations. Compared to the control group, superoxide dismutase and glutathione peroxidase activities in the photoaging group were significantly reduced, while malondialdehyde and reactive oxygen species content, along with tumor necrosis factor-alpha (TNF-a) and interleukin (IL)-10 mRNA and protein levels were markedly increased. In contrast, Bcl-2 protein expression was significantly decreased, while caspase-3, caspase-9, and Bax protein expression levels were significantly increased. Compared to the photoaging group, HaCaT cell proliferation was significantly increased in the PE group. Both PE concentrations significantly increased superoxide dismutase and glutathione peroxidase activities in cells, reduced malondialdehyde and reactive oxygen species content, decreased TNF-a and IL-10 mRNA expression in cells, and reduced TNF-a and IL-10 protein levels in the supernatant. Additionally, Bcl-2 protein expression levels were significantly increased, while caspase-3, caspase-9, and Bax protein expression levels were significantly reduced by PE treatment. Conclusions: PE can inhibit UV-induced apoptosis by inhibiting mitochondria-mediated apoptosis and regulating TNF-a and IL-10 expression.


Author(s):  
Mingyang Li ◽  
Chunli Qi ◽  
Renxing Song ◽  
Chunming Xiong ◽  
Xiao Zhong ◽  
...  

Cardiac fibrosis is a hallmark of various heart diseases and ultimately leads to heart failure. Although long noncoding RNA (lncRNA) SNHG20 has been reported to play important roles in various cancers, its function in cardiac fibrosis remains unclear. The expression of SNHG20 and miR-335 in heart tissues of Angiotensin II induced mice and Angiotensin II stimulated mouse cardiomyocyte cell line HL-1 were detected by qRT-PCR. Cell viability was evaluated by CCK-8 assay. The expression of Galectin-3, fibrosis-related proteins (Fibronectin, Collagen IaI and α-SMA) and apoptosis-related proteins (Cleaved Caspase-3 and Cleaved PARP) were detected by Western blot. Bioinformatics prediction, luciferase reporter assay and RNA pull down assay were performed to determine the relationship between SNHG20 and miR-335, as well as miR-335 and Galectin-3 . Gain- and loss-function assays were performed to determine the role of SNHG20 /miR-335/ Galectin-3 in cardiac fibrosis. SNHG20 was significantly upregulated, and miR-335 was downregulated in heart tissues of Angiotensin II treated mice and Angiotensin II stimulated HL-1 cells. Downregulation of SNHG20 effectively enhanced cell viability, while decreased cell size of HL-1 cells and the expression levels of fibrosis-related proteins (Fibronectin, Collagen IaI and α-SMA) and apoptosis-related proteins (Cleaved Caspase-3 and Cleaved PARP), which was induced by Angiotensin II treatment. Furthermore, SNHG20 elevated the expression levels of Galectin-3 by directly regulating miR-335. Our study revealed that downregulation of SNHG20 improved Angiotensin II-induced cardiac fibrosis by targeting the miR-335/ Galectin-3 axis, suggesting that SNHG20 may be a potential therapeutic target for cardiac fibrosis and hypertrophy.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8562 ◽  
Author(s):  
Haibo Yao ◽  
Fang Lu ◽  
Yanfei Shao

Background The E2F family is a group of genes encoding a series of transcription factors in higher eukaryotes and participating in the regulation of cell cycle and DNA synthesis in mammals. This study was designed to investigate the role of E2F family in colon cancer. Methods In this study, the transcriptional levels of E2F1-8 in patients with colon cancer from GEPIA was examined. Meanwhile, the immunohistochemical data of the eight genes were also obtained in the The Human Protein Atlas website. Additionally, we re-identified the mRNA expression levels of these genes via real time PCR. Furthermore, the association between the levels of E2F family and stage plot as wells overall survival of patients with colon cancer were analyzed. Results We found that the mRNA and protein levels of E2F1, E2F2, E3F3, E2F5, E2F7 and E2F8 were significantly higher in colon cancer tissues than in normal colon tissues while the expression levels of E2F4 and E2F6 displayed no significant difference between colon cancer tissues and normal tissues. Additionally, E2F3, E2F4, E2F7 and E2F8 were significantly associated with the stages of colon cancer. The Kaplan-Meier Plotter showed that the high levels of E2F3 conferred a worse overall survival and disease free survival of patients with colon cancer. Also, high levels of E2F4 resulted in a worse overall survival. Conclusion Our study implied that E2F3, E2F4, E2F7 and E2F8 are potential targets of precision therapy for patients with colon cancer while E2F1, E2F2, E3F3, E2F5, E2F7 and E2F8 are potential biomarkers for the diagnosis of colon cancer.


2020 ◽  
Vol 14 (4) ◽  
pp. 579-583
Author(s):  
Tian-Li Chen ◽  
Tong-Tong Song ◽  
Xu Sun ◽  
Yi-Wen Gu ◽  
Wen-Jing Du ◽  
...  

To investigate the protective effect of Angelica sinensis polysaccharide (ASP) on spleen injury and its influence of Bcl-2/Bax/Caspase-3 signal pathway in radiation rats, 60 Sprague Dawley (SD) rats were divided into 5 groups randomly: the normal group, the model group, low-, middle- and high- dose of ASP groups. On the 15th day after the appropriate medication, all the rats except the normal group received the 60Co γ-ray irradiation once with a total dose of 4.0Gy. The levels of NF-kBp65 and IKKa in serum, the contents of MDA, GSH and SOD in spleen of each group were detected with ELISA after 24 h of irradiation, and the protein expression levels of Bcl-2, Bax, Caspase-3, HSPBP1 and TIMM8B in spleen of each group were detected by western blotting. The results showed that compared with the model group, NF-kBp65, IKKa and MDA contents were decreased while GSH and SOD contents were increased in the middle and high dose groups. The level of Bcl-2 and TIMM8B were increased while Bax, caspase-3 and HSPBP1 were decreased in all groups. In conclusion, Angelica sinensis polysaccharide can reduce the expression levels of Bax and caspase-3, while increase the level of Bcl-2 in spleen of rats induced by radiation and antagonize the body injury caused by radiation by regulating Bcl 2/Bax/Caspase 3 signaling pathway.


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